7196 research outputs found
Sort by
Timut Pepper Extract Slows Age-Dependent Decline of Mobility and Collagen Loss and Promotes Longevity.
No full textInvestigations into human longevity are increasingly focusing on healthspan enhancement, not just lifespan extension. Lifestyle modifications and nutritional choices, including food supplements, can significantly affect aging and general health. Phytochemicals in centenarians' diets, such as those found in Timut pepper, a Nepalese spice with various medicinal properties, may contribute to their longevity. Similarly, Sichuan pepper, a related species, has demonstrated anti-inflammatory and neuroprotective activities. With the broader purpose of uncovering a novel treatment to address aging and its comorbidities, this study aims to investigate the potential lifespan- and healthspan-promoting effects of Timut pepper using the model organism We show that Timut pepper extract extends lifespan at different maintenance temperatures and increases the proportion of active nematodes in their early adulthood. In addition, we show that Timut pepper extract enhances speed and distance moved as the nematodes age. Finally, Timut pepper extract assures extracellular matrix homeostasis by slowing the age-dependent decline of collagen expression
An Additional Lrp4 High Bone Mass Mutation Mitigates the Sost-Knockout Phenotype in Mice by Increasing Bone Remodeling.
No full textPathogenic variants disrupting the binding between sclerostin (encoded by SOST) and its receptor LRP4 have previously been described to cause sclerosteosis, a rare high bone mass disorder. The sclerostin-LRP4 complex inhibits canonical WNT signaling, a key pathway regulating osteoblastic bone formation and a promising therapeutic target for common bone disorders, such as osteoporosis. In the current study, we crossed mice deficient for Sost (Sost-/-) with our p.Arg1170Gln Lrp4 knock-in (Lrp4KI/KI) mouse model to create double mutant Sost-/-;Lrp4KI/KI mice. We compared the phenotype of Sost-/- mice with that of Sost-/-;Lrp4KI/KI mice, to investigate a possible synergistic effect of the disease-causing p.Arg1170Trp variant in Lrp4 on Sost deficiency. Interestingly, presence of Lrp4KI alleles partially mitigated the Sost-/- phenotype. Cellular and dynamic histomorphometry did not reveal mechanistic insights into the observed phenotypic differences. We therefore determined the molecular effect of the Lrp4KI allele by performing bulk RNA sequencing on Lrp4KI/KI primary osteoblasts. Unexpectedly, mostly genes related to bone resorption or remodeling (Acp5, Rankl, Mmp9) were upregulated in Lrp4KI/KI primary osteoblasts. Verification of these markers in Lrp4KI/KI, Sost-/- and Sost-/-;Lrp4KI/KI mice revealed that sclerostin deficiency counteracts this Lrp4KI/KI effect in Sost-/-;Lrp4KI/KI mice. We therefore hypothesize that models with two inactivating Lrp4KI alleles rather activate bone remodeling, with a net gain in bone mass, whereas sclerostin deficiency has more robust anabolic effects on bone formation. Moreover, these effects of sclerostin and Lrp4 are stronger in female mice, contributing to a more severe phenotype than in males and more detectable phenotypic differences among different genotypes
The title was changed to "Implementation of micelle-enabled C(sp2)–C(sp3) cross-electrophile coupling in pharmaceutical synthesis" during revision
No full textA sustainable C(sp2)-C(sp3) cross- electrophile coupling was developed between readily available 5-bromophthalide and 1-benzyl-4-iodopiperidine under micellar conditions in water, leading to the key intermediate of one of our development compounds. Copper was found to play a crucial role as co-catalyst in this due catalysis system. The chemistry and process were successfully demonstrated in kilo scale to deliver sufficient drug substance to the clinical campaigns
Automated high throughput workflow for rapid implementation of immobilized enzymes in chemical process development
No full textDespite known advantages of immobilized enzymes, their broad application in pharmaceutical drug manufacturing has not yet caught up. Limited access to commercially available immobilized enzymes on one side and time - and cost - consuming development of new processes on the other side have hampered their broader implementation so far. Herein, we present a walk-away high throughput workflow with state of the art robotic equipment that enables the rapid optimization and development of immobilized enzymes - ready to be scaled - and thus matching the need for speed in modern drug development. Following this workflow we were able to identify an engineered ketoreductase which was >2-fold more active than the wildtype from L. kefir. Further optimzation of the reaction and immobilization conditions lead to an additional 29-fold productivitiy increase from 0.36 to 10.6 gproduct/genzyme for the selected enzyme variant
Capmatinib plus nivolumab in pretreated patients with EGFR wild-type advanced non-small cell lung cancer.
No full textDysregulated MET is an established oncogenic driver in non-small cell lung cancer (NSCLC). MET signaling may also suppress anticancer immune responses. Concomitant MET inhibition with capmatinib (a MET inhibitor) synergistically enhanced the efficacy of immunotherapies in murine cancer models, regardless of tumor dependency to MET signaling. Here, we report results of a multicenter, open-label, phase 2 study of capmatinib plus nivolumab (a PD-1 inhibitor) in patients with EGFR wild-type advanced NSCLC, previously treated with platinum-based chemotherapy.Patients were allocated into high-MET or low-MET groups according to MET expression determined by immunohistochemistry, MET gene copy number as assessed by fluorescence in-situ hybridization, and presence of MET exon 14 skipping mutation, then received capmatinib 400 mg, oral, twice daily in combination with nivolumab 3 mg/kg intravenously every 2 weeks. The primary endpoint was investigator-assessed 6-month progression-free survival (PFS) rate per RECIST v1.1.The primary endpoint was met in both the high-MET (N = 16) and low-MET (N = 30) groups. In the high-MET and low-MET groups, respectively, the estimated mean 6-month PFS rate (95 % credible interval) by Bayesian analysis was 68.9 % (48.5-85.7) and 50.9 % (35.6-66.4). The Kaplan-Meier median PFS (95 % CI) was 6.2 months (3.5-19.2) and 4.2 months (1.8-7.4). The overall response rate (95 % CI) was 25.0 % (7.3-52.4) and 16.7 % (5.6-34.7). Most frequent treatment-related adverse events (≥30 % any grade, N = 46) were nausea (52.2 %), peripheral edema (34.8 %), and increased blood creatinine (30.4 %).Capmatinib plus nivolumab showed clinical activity and manageable safety in pretreated patients with advanced EGFR wild-type NSCLC, independent of MET status.ClinicalTrials.gov NCT02323126
Position Statement in Support of Cell Bank Stability Monitoring from a Single Storage Location of Equivalent Storage Condition
No full textEvidence of stability of cell banks, used as a starting material for medicinal biotech products, under defined conditions of long-term storage is one requirement to ensure cell substrate stability per ICHQ5D and WHO TRS 978 Annex 3. Storing cell banks at appropriate storage conditions serves to halt molecular movement and biological activity, thus maintaining the cell banks' stability attributes, such as stable cell viability and growth upon reconstitution.
A stability monitoring program verifies this and ensures continuous quality oversight.
Stability monitoring programs are based on pre-defined bank-specific criteria. Representative data including all stability relevant parameters for a Master Cell Bank (MCB) are generated by initial characterization, production of clinical or commercial material or preparation of a new Working Cell Bank (WCB). Belonging data for WCB are generated from thaws for a clinical or commercial production campaign or appropriately documented scale-down or pilot-scale processes. Cell bank stability can also be monitored at a pre-defined interval in case no suitable thaws take place.
In alignment with ICHQ5D, redundancy in the storage of cell bank containers applies for MCB and WCB storage to provide protection from catastrophic events and to ensure continuous, uninterrupted production of pharmaceuticals. Common redundant storage strategies include storing portions of a single cell bank in different freezers, various physical locations within one site, or storing a portion of the cell bank at a redundant site.
It is common practice across the industry not to include the evaluation of cell bank vials stored at all redundant storage sites as part of the stability monitoring program. Redundant storage sites abiding by the same storage condition requirements and equivalent quality management system are equivalent in function, ensuring comparable and stable quality of cell banks across all sites.
This approach is justified by the following rationales:
Storage equipment is qualified, monitored and suitable for cell bank storage at the appropriately defined temperatures for the respective cell bank type. Operational parameters like temperature ranges and alarm limits are in place according to cell bank storage specifications. At all locations, operation and maintenance of storage equipment follow defined procedures, leading to an equivalent operational status of the equipment. Inventory oversight, routine documentation of withdrawals and entries as well as access control are in place at each redundant site according to the GMP standard for maintaining the cell banks' quality attributes.
Cell bank transportation for long-term storage takes place in qualified shipping containers and by qualified delivery companies under the same conditions as long-term storage. Transportation controls are in place to ensure temperature is maintained within specification throughout the transfer. Excursions of storage and shipment temperature, which might occur at all redundant storage locations, are assessed in the quality system for product quality impact, root causes and potential corrective actions.
In summary, incorporating stability monitoring for all redundant storage sites will lead to quicker depletion of cell banks which are intended to support production of clinical and commercial material, while generating data of no additional informative value to evaluate the storage stability of a cell bank. Under equivalent storage and quality management procedures, the quality of cell banks stored at redundant locations should be considered comparable due to the aforementioned reasons
Sequestration of membrane cholesterol by cholesterol-binding proteins inhibits SARS-CoV-2 entry into Vero E6 cells
No full textMembrane lipids and proteins form dynamic domains crucial for physiological and pathophysiological processes, including viral infection. Many plasma membrane proteins, residing within membrane domains enriched with cholesterol (CHOL) and sphingomyelin (SM), serve as receptors for attachment and entry of viruses into the host cell. Among these, human coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), use proteins associated with membrane domains for initial binding and internalization. We hypothesized that the interaction of lipid-binding proteins with CHOL in plasma membrane could sequestrate lipids and thus affect the efficiency of virus entry into host cells, preventing the initial steps of viral infection. We have prepared CHOL-binding proteins with high affinities for lipids in the plasma membrane of mammalian cells. Binding of the perfringolysin O domain four (D4) and its variant D4E458L to membrane CHOL impaired the internalization of the receptor-binding domain of the SARS-CoV-2 spike protein and the pseudovirus complemented with the SARS-CoV-2 spike protein. SARS-CoV-2 replication in Vero E6 cells was also decreased. Overall, our results demonstrate that the integrity of CHOL-rich membrane domains and the accessibility of CHOL in the membrane play an essential role in SARS-CoV-2 cell entry
The Future Is Not Bright: Evaluation of Rat Preferences for Color and Intensity of Light
No full textLight is a key factor influencing the welfare of laboratory rodents, but little is known about their optimal lighting condition. It i common knowledge that rats prefer dim light, so bright light is mitigated with red-tinted shelters or cages, which alter both the color and intensity of light. Because both aspects are altered, the contribution of each feature to rodent preference is unknown. Further, it is unknown if this preference is influenced by previous experience. We hypothesized that rats would prefer lower light intensity and that their preferences would be influenced by their housing environment. Breeder pairs of rats were randomly separated into four treatments groups: red 200 lux, red 25 lux, clear 200 lux, and clear 25 lux. The breeders’ offspring were tested three times in an apparatus that offered access to each environment, and their preferences were analyzed. Generally, the rats preferred the lower-lux environments and showed no color preference. However, the rats from the clear, 200 lux cages, preferred clear caging and only showed a preference for 25 lux conditions during the second and third preference tests. These results suggest that the light intensity, more than color, should be considered when designing rodent housing and testing facilities
EVALUATION OF THE EMA TRIGGER FOR FISH BCF TESTING AND SECONDARY POISONING ASSESSMENT: EVALUATION OF LOG D AND FISH BCF DATA FOR SEVERAL PHARMACEUTICALS
No full textAs per the European Medicines Agency (EMA) Guideline for Environmental Risk Assessment of Medicinal Products for Human Use (EMA 2006), a fish bioconcentration study is triggered in Phase I for pharmaceuticals having a log Kow >4.5 to support screening for Persistence, Bioaccumulation and Toxicity (PBT) and in Phase II for pharmaceuticals having a log Kow >3 to assess bioaccumulation potential. The recommended protocol for bioaccumulation is OECD Test Guideline 305: Bioaccumulation in Fish, Aqueous and Dietary Exposure (OECD 2012). Based on the standard OECD 305 sampling schedule, approximately 200 fish per study may be required to determine steady-state (BCFSS) and kinetic (BCFK) bioconcentration factors following fish exposures to a low- and high-test concentration and a negative control. As per the draft revision of the EMA guideline, released for consultation in 2018, when log Kow is ≥3 the potential for secondary poisoning should be evaluated by first determining the experimental fish BCF, and when the resulting BCF is >100 L kg-1, a secondary poisoning assessment is required. Considering the potential for fish to metabolize and excrete xenobiotics and the number of animals required for a standard BCF test, the BCF values for several pharmaceuticals were evaluated to understand whether existing data support the current log Kow trigger of 3 for BCF testing. In the interest of animal welfare, BCF data were also evaluated to determine if study design could be limited to a single test concentration and still provide a good estimate of the BCF while reducing the number of fish required. Based on the data presented, increasing the log Kow trigger for BCF testing from 3 to 4 is proposed, and use of a single test concentration to evaluate bioaccumulation potential is recommended. Future plans include an evaluation of the proposed EMA BCF trigger for a secondary poisoning assessment